脾氨肽口服溶液对大鼠变应性鼻炎复发的疗效及作用机制研究

Study on the therapeutic effect and mechanism of spleen aminopeptide oral solution on the recurrence of allergic rhinitis in rat

目的构建变应性鼻炎(allergic rhinitis,AR)复发大鼠模型,探究脾氨肽口服溶液防治AR复发的疗效及作用机制。方法32只健康SPF级SD雌性大鼠按随机数字表分为空白组、模型组、脾氨肽组、氯雷他定组,每组8只。采用卵清蛋白结合氢氧化铝佐剂诱导AR复发大鼠模型。造模成功后,空白组、模型组不给药,脾氨肽组予脾氨肽口服溶液(7.56 mg/kg),氯雷他定组予口服氯雷他定(1.26 mg/kg),之后进行二次激发。鼻部症状行为学评分评价各组大鼠治疗前后症状改善情况。血常规检测观察各组大鼠外周血嗜酸性粒细胞、嗜碱性粒细胞等水平。苏木精—伊红染色观察鼻甲组织病理形态。Elisa法检测各组大鼠血清抗体水平。流式细胞检测各组大鼠外周血免疫细胞及炎症因子水平。结果(1)鼻部症状行为学评分:模型组评分总体处于最高水平,脾氨肽组和氯雷他定组均低于模型组;(2)嗜酸性粒细胞数量和嗜碱性粒细胞数量:与模型组相比,脾氨肽组和氯雷他定组嗜酸性细胞数量和嗜碱性粒细胞数量均降低;(3)鼻组织病理观察:脾氨肽组鼻黏膜病变轻于模型组和氯雷他定组;(4)Elisa检测:与模型组相比,脾氨肽组和氯雷他定组IgG和卵清蛋白特异性IgG的OD值均升高;(5)流式检测结果:与模型组相比,脾氨肽组、氯雷他定组辅助性T细胞1(T helper 1 cell,Th1)和Treg细胞均升高,Th2和Th17细胞均降低,且脾氨肽组Th2细胞下降明显(P<0.05);(6)炎症因子检测结果:与模型组相比,脾氨肽组γ干扰素、肿瘤坏死因子-α、粒细胞巨噬细胞集落刺激因子等分泌增多,白介素4、白介素5、白介素6等分泌减少。结论脾氨肽口服溶液能有效减轻AR复发鼻部症状,缓解鼻部炎症反应,修复鼻黏膜损伤。脾氨肽口服溶液能抑制白介素4分泌,降低AR复发大鼠Th2比例,同时促进γ干扰素分泌,提高Th1比例,增强单核巨噬系统活性,激活细胞免疫,调节免疫失衡,从而起到防治AR复发的作用。

Objective To explore the efficacy and mechanism of Splenic Aminopeptide oral solution in the treatment of AR recurrence by constructing a rat model of AR recurrence.Methods The rat model of AR recurrence was induced by ovalbumin(OVA)combined with aluminum hydroxide adjuvant,and loratadine was used as a positive control drug.The therapeutic effect of oral splenic peptide solution to prevent AR recurrence was explored by nasal symptom behavioral score,blood routine test,and nasal mucosa pathological tissue test.The mechanism of oral splenic peptide solution to prevent AR recurrence was explored by Elisa test,flow cytometry,and multifactor test.Results(1)Behavioral score of nasal symptoms:The score of the model group was generally at the level,both in the spleen aminopeptide group and the loratadine group were lower than that in the model group;(2)The number of eosinophil and basophil:Compared with the model group,the number of eosinophil and basophil in spleen amine peptide group and loratadine group were all;(3)Pathological observation of nasal tissue:The lesions of nasal mucosa in the splenaminopeptide group were lighter than that in the model group and the loratadine group;(4)Elisa:Compared with the model group,the levels of IgG and OVA-specific IgG OD in the spleen aminopeptide group and the loratadine group were increased;(5)Flow detection results:Compared with the model group,the levels of Th1 and Treg in the spleen aminopeptide group were increased.The levels of Th2 and Th17 were decreased,while Th2 in the spleen aminopeptide group was decreased significantly(P<0.05);(6)Inflammatory factors:IFN-γ,TNF-α,GM-CSF increased,IL-4,IL-5,IL-6 in the splenaminopeptide group compared with model group.Conclusions Spleen aminopeptide oral solution can effectively alleviate the nasal symptoms of AR recurrence,alleviate the nasal inflammatory reaction,and repair the nasal mucosa damage.The mechanism of spleen aminopeptide oral solution in preventing and treating AR recurrence is to reduce the level of Th2 cells in rats with AR recurrence,significantly reduce the level of IL-4,inhibit humoral immunity,increase the proportion of Th1 cells,and stimulate INF-γ.It can secrete,enhance the activity of mononuclear macrophage system,activate cellular immunity,and regulate immune imbalance.